Antibiosis of the pith maize to Sesamia nonagrioides (Lepidoptera: Noctuidae)

Thirteen inbred lines of maize (Zea mays L.) with different levels of stem resistance to the stem borer Sesamia nonagrioides Lefevbre were evaluated in the field and the laboratory to determine the antibiotic resistance to this pest. Inbreds CM151, CO125, and EP39 had antibiotic pith as well as stem resistance, so the pith could play a role in stem resistance. Inbreds A509, F473, and PB130 did not have antibiotic pith but had stem resistance; therefore, other mechanisms could confer stem resistance. Finally, the inbred MS1334 had antibiotic pith and did not show stem resistance; thus, other factors could compensate the effect of the pith. Therefore, although pith antibiotic compound seems to play a role in the defense against S. nonagrioides attack, it is not the only possible mechanism of defense.     

Performance of crosses among flint maize populations under infestation by Sesamia nonagrioides (Lepidoptera: Noctuidae)

Flint maize, Zea mays L., varieties provide some interesting agronomic characteristics and kernels that possess a better ability than other kernels for developing high-quality flour. The pink stem borer, Sesamia nonagrioides Lefebvre, is an important constraint for the maize crop in Mediterranean regions. The objective of this work was to identify a "flint x flint" heterotic pattern that would perform well under artificial infestation by S. nonagrioides. A 10-population diallel was evaluated under infestation by S. nonagrioides in 2 yr. Variety effects were the only significant effects involved in stem and ear resistance to S. nonagrioides attack. Variety effects and average heterosis effects were the only significant effects for grain yield under artificial infestation conditions. Considering variety effects and cross-performance, the heterotic pattern Basto/Enano levantixo x Longfellow (BA/EL x LO) would be recommended for obtaining flint maize hybrids tolerant to S. nonagrioides attack because BA/EL had the most favorable variety effects for stem resistance, LO exhibited the most positive variety effects for grain yield, and the cross BA/EL x LO yielded significantly more than the remaining crosses.     

Genetic variation among Mediterranean populations of Sesamia nonagrioides (Lepidoptera: Noctuidae) as revealed by RFLP mtDNA analysis

Restriction fragment length polymorphism analysis of two segments of mitochondrial DNA (COI and 16S rRNA) was used to examine genetic variation in Sesamia nonagrioides (Lefèbvre) populations from the Mediterranean basin. Four populations were collected from central and southern Greece, and five from northern latitudes: Greece, Italy, France and Spain. No variation was observed in COI, while 16S rRNA segment proved highly polymorphic and 28 different haplotypes were found. Lower intra-population polymorphism was found in the northern populations than in southern ones. Although no significant isolation by distance was found, the UPGMA tree based on Nei's raw number of nucleotide differences separated the populations into two major groups, i.e. one with the northern (40.6 degrees N-43.4 degrees N) and the other with the southern populations (37.3 degrees N-39.2 degrees N). Analysis of molecular variance revealed that most of the variation was between the two major groups (Phi(CT)=0.559), and all pairwise comparisons between the northern and southern populations resulted in high and significant F(ST) values (overall F(ST)=0.604). The high F(ST) values and the strong spatial genetic structure indicate that long-distance migration may be a rare event. The populations do not seem to have experienced a strong historical bottleneck. The occurrence of a few widespread haplotypes and the genetic similarity of the northern populations could be attributed to a historical expansion of certain haplotypes from the south towards to the northern borders of the species' distribution area.     

Inheritance of resistance to ear damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae) in maize

Sesamia nonagrioides Lefebvre is a major insect pest of maize (Zea mays L.) in northwestern Spain. The inheritance of ear resistance in field corn to this pest has not been studied. This work aims to determine the importance of genetic (additive, dominance, and epistatic effects) and environmental effects in the inheritance of ear resistance to S. nonagrioides. Three field corn inbreds (CM109, EP31, and EP42) were used as parents and two crosses (CM109 x EP31 and CM109 x EP42) were made. These inbreds show different ear resistance levels to S. nonagrioides, with CM109 more resistant than EP31 and EP42. For each cross, parents (P1, P2), F1, F2, and backcrosses to each parent (BC1 and BC2) were evaluated. Correlations among ear damage traits showed that general appearance of the ear should be useful indicator of ear resistance. Ear resistance was dominant to susceptibility and was transmitted from inbreds to their hybrids. The additive-dominance model fit the generation mean analysis for both crosses and the degree of genetic control varied depending on the cross and trait. For both crosses, additive and dominance effects were significant for most ear damage traits. Epistatic gene effects were significant for husk and shank damage, and gene effects for number and length of tunnels were not significant. Because ear resistance involved additive and dominance effects for this set of inbreds, breeding procedures based on both types of gene action should be effective.     

Fine structure of the female sex pheromone-producing glands in Sesamia nonagrioides lef. (Lepidoptera : Noctuidae)

The morphology and cytology of the female sex pheromone-producing glands of Sesamia nonagrioides (Lepidoptera : Noctuidae) consist of modified intersegmental membranes; they are unusual compared with those described in other moths. There are 2 different glandular areas (a ventral area, GI, around the ostium bursae and a ring-shaped area, GII, around the ovipositor), instead of a single area (between segments 8 and 9 + 10) as in most other species. Ultrastructural features, such as developed basal invaginations and dilated intercellular spaces indicate transport of material from the hemolymph to the glandular cells, while the presence of abundant mitochondria, numerous free ribosomes, rough endoplasmic reticulum, and inclusions reveal high cellular activity. The smooth endoplasmic reticulum is absent in S. nonagrioides, in contrast to its extensive development in glands producing similar compounds in other moths. The cuticle, abnormally thick (7–8 times thicker than the glandular epithelium), is traversed by dilated pore canals forming cuticular gaps, connected with epicuticular canals. This system allows the storage and the passage of pheromone in and through the cuticle.     

Resistance of early maturing sweet corn varieties to damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae)

In Mediterranean countries, the principal pest of maize, Zea mays L., is Sesamia nonagrioides Lefebvre. The objective of this work was to study the resistance of the four early maturing varieties of sweet corn, Baby Orchard, Orchard Baby, Dorinny Sweet, and Golden Early Market, and to check the relationship among resistance to S. nonagrioides and infestation dates. In a previous study, these varieties had showed a confusing behavior, being the most resistant varieties to S. nonagrioides attack in some environments and the most susceptible in others. These varieties were again evaluated along with three medium maturing varieties, used as testers because they were more stable in both environments, by using two infestation dates. Plants were more damaged in the late infestation, but the effect of the infestation depends on the variety studied. Orchard Baby and Baby Orchard were the most resistant varieties under two infestation dates. These two varieties are closely related. Therefore, we can use either one as a source of precocity and resistance for modern sweet corn. Although many studies showed that early maturing inbreds had greater damage caused by S. nonagrioides and European corn borer than late maturing inbreds and hybrids, our results showed that the earliest maturing varieties were the most resistant.     

Chemical stimuli from corn plants affect host selection and oviposition behavior of Sesamia nonagrioides (Lepidoptera: Noctuidae)

The role of chemical stimuli in host selection and oviposition of Sesamia nonagrioides (Lef) was studied in the laboratory using eight corn hybrids. Female oviposition preference was evaluated on plants of the eight hybrids, their extracts in pentane and methanol, and volatiles collected by steam distillation in ether. Results from two-choice bioassays indicate various degrees of female preference to the hybrids. Four hybrids received more eggs than control plants, whereas the remainder received fewer eggs. Relatively similar results were obtained when filter paper sticks baited with pentane and steam distillation extracts were evaluated. Methanol extracts from all eight hybrids received fewer eggs than the methanol controls. Results showed that pentane and steam distillation extracts contain chemicals that elicit female oviposition response, while methanol extracts may contain chemicals that deter or repel females to oviposition.     

Free phenols in maize pith and their relationship with resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) attack

The stem borer Sesamia nonagrioides (Lefèbvre) is the most important insect pest of maize, Zea mays L., in northwestern Spain. Among the metabolites present in maize, phenolic compounds could play an important role in resistance. The objective of this work was to determine whether a relationship between phenols and the amount of resistance exists. Amounts of free phenolic compounds in the pith of 13 inbred maize lines that differ in resistance were measured. The phenolic compounds identified were p-coumaric acid, cafeic acid, ferulic acid, vanillic acid, syringic acid, chorogenic acid, sinapic acid, p-hydroxybenzoic acid, and vanillin. The amount of free p-coumaric acid was correlated with the resistance level. Higher quantities of p-coumaric in the pith could contribute to general resistance to stem borer attack. Jointly with ferulic acid, p-coumaric could provide resistance mechanisms through cell wall fortification and lignification. The other compounds showed no or an unclear relationship with resistance. The vanillic acid showed a decreased tendency after silking, when maize is most attractive for S. nonagrioides, suggesting this acid could act as a chemoattractant for S. nonagrioides larvae or adults. Future studies that focus on these phenolic compounds could be useful in understanding S. nonagrioides resistance.     

Mean generation analysis of the damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) in sweet corn ears

Sesamia nonagrioides (Lefebvre) and Ostrinia nubilalis (Hübner) are the main maize (Zea mays L.) pests in Mediterranean countries. To develop insect-resistant cultivars, it is helpful to know the genetic control of the resistance. Our objective was to determine the genetic control of the resistance to both borers. For each of two crosses (EP59 x P51 and 15125 x EP61), six generations were evaluated (P1, P2, F1, F2, BC1, and BC2). Genetic effects x environment interactions were not significant. For the O. nubilalis resistance traits; both crosses fitted an additive-dominance model. EP59 x P51 had important dominance and additive effects, whereas for 15125 x EP61 we did not detect significant genetic effects, but significant year effects were detected. For S. nonagrioides infestation, both crosses fitted to an additive-dominance model. There were additive effects for most traits in EP59 x P51. The cross I5125 x EP61 showed significant dominance effects for several traits. Resistance to both corn borers fit an additive-dominance model, but genetic effects depend on the cross evaluated. In the resistance to S. nonagrioides, additive effects were important for shank resistance, which is a useful trait for avoiding S. nonagrioides damage on the ear. Early sowings are recommended to make good use of the resistance to both corn borers. In the late sowings, damage is so high that resistant plants are not able to control the pest.     

Evaluation of the European Union Maize Landrace Core Collection for resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae)

Two corn borer species are the principal maize insect pests in Europe, the European corn borer, Ostrinia nubilalis (Hübner), and the pink stem borer, Sesamia nonagrioides (Lefebvre). Hence, it would be advisable to evaluate the European maize germplasm for corn borer resistance to generate European varieties resistant to corn borer attack. The creation of the European Union Maize Landrace Core Collection (EUMLCC) allowed the screening of most of the variability for European corn borer resistance present among European maize local populations from France, Germany, Greece, Italy, Portugal, and Spain, testing a representative sample. The objective of this study was the evaluation of stem and ear resistance of the EUMLCC to European corn borer and pink stem borer attack. Trials were made at two Spanish locations that represent two very different maize-growing areas. Populations that performed relatively well under corn borer infestation for stem and ear damage were 'PRT0010008' and'GRC0010085', among very early landraces; 'PRT00100120' and 'PRT00100186', among early landraces; 'GRC0010174', among midseason landraces; and 'ESP0070441', among late landraces. Either the selection that could have happen under high insect pressure or the singular origin of determined maize populations would be possible explanations for the higher corn borer resistance of some landraces. Landraces 'PRT0010008', 'FRA0410090', 'PRT00100186', and 'ESP0090214' would be selected to constitute a composite population resistant to corn borers and adapted to short season, whereas populations 'ESP0090033', 'PRT00100530', 'GRC0010174', and 'ITA0370005' would be used to make a resistant composite adapted to longer season.     

Characterization and distribution of digestive proteases of the stalk corn borer,Sesamia nonagrioides Lef. (Lepidoptera: Noctuidae)

Larval midgut extracts from the noctuid Sesamia nonagrioides Lef. were assayed for protease activity. Total proteolytic activity, as measured by azocasein hydrolysis, showed a pH optimum in the range 10.0 to 11.5, suggesting a digestive system based largely on serine-like proteases. The ability of midgut extracts to hydrolyze specific synthetic substrates, the elucidation of the pH at which maximal hydrolysis occurs, and their sensitivity to protease inhibitors confirmed the presence of the serine endoproteases: trypsin, chymotrypsin, and elastase; and the exopeptidases: carboxypeptidase A, carboxypeptidase B, and leucine aminopeptidase. The distribution of these digestive proteases along the gut sections and among the different midgut regions was examined. All types of endoproteases and exopeptidases were mainly located in the midgut, with less than 5% of the activity in the foregut and hindgut. When the two halves of the midgut were compared, all proteolytic activities were higher in the anterior portion of the midgut. Trypsin, chymotrypsin, elastase, and carboxypeptidase B activities were mainly located in the endoperitrophic space of the midgut, with some activity in the ectoperitrophic space, whereas aminopeptidase and carboxypeptidase A activities were preferentially located in the midgut epithelium. © 1996 Wiley-Liss, Inc.     

Susceptibility of Spanish populations of the corn borers Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) to a Bacillus thuringiensis endotoxin

Baseline susceptibility to the Cry1Ab delta-endotoxin from Bacillus thuringiensis (Berliner) was determined for four populations of Sesamia nonagrioides (Lefebvre) and two populations of Ostrinia nubilalis (Hübner) from Spain. This study shows that S. nonagrioides is at least as susceptible as O. nubilalis to B. thuringiensis Cry1Ab protein. We found small differences in susceptibility among the Spanish populations of S. nonagrioides that can be attributed to natural variation, because there are no records of B. thuringiensis products being used on corn crops in Spain. There were no differences in susceptibility to Cry1Ab toxin between the two populations of O. nubilalis.     

Frequency of resistance to Bacillus thuringiensis toxin CrylAb in Greek and Spanish population of Sesamia nonagrioides (Lepidoptera: Noctuidae)

The high-dose/refuge strategy is considered as the main strategy for delaying resistance in target pests to genetically modified crops that produce insecticidal proteins derived from Bacillus thuringiensis Berliner. This strategy is based on a key assumption that resistance alleles are initially rare (<10(-3)). To test this assumption, we used an F2 screen on natural populations of Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae) from Greece and Spain. In total, 75 lines from Greece and 85 lines from Spain were screened for survival of F2 larvae on Cry1Ab corn, Zea mays L., leaves. No major resistance alleles were found. The frequency of resistance alleles in the Greek population was <9.7 x 10(-3) with 95% probability, which was very similar to that of the Spanish population (<8.6 x 10(-3) with 95% probability), and the expected frequencies were 3.2 x 10(-3) (0-0.0097) and 2.9 x 10(-3) (0-0.0086) in Greece and Spain (pooled 1.5 x 10(-3)). The experiment-wise detection probability of resistance was 94.0 and 97.5% for the Greek and the Spanish population, respectively. Evidence of alleles conferring partial resistance to Cry1Ab was found only for the Greek population. The frequency of alleles for partial resistance was estimated as 6.5 x 10(-3) with a 95% credibility interval between 8 x 10(-4) and 17.8 x 10(-3) and a detection probability of 94%. Our results suggest that the frequency of alleles conferring resistance to CrylAb, regarding the population of S. nonagrioides, may be rare enough so that the high-dose/refuge strategy could be applied with success for resistance management.     

Factors affecting the natural duration of diapause and post-diapause development in the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae)

Sesamia nonagrioides responds to photoperiod for diapause termination at high temperature with a Type II response curve, and only photoperiods longer than 12:12 terminate diapause. However, these photoperiods never occur in the field when diapausing larvae are competent to terminate diapause. Under a temperature similar to the natural field temperatures diapause terminates spontaneously in approximately 4 months, which ensures that the larvae reach the middle of winter without pupation. S. nonagrioides larvae pupate after going through a specific number of light-dark cycles or days: the required day number (RDN) for diapause completion. This RDN could be modulated by temperature but more research is necessary to clarify this point. In post-diapause development, when a suitable temperature threshold is considered, the absolute accumulation of heat is more important than whether the temperature received is fluctuating or constant. The temperature threshold for diapause and post-diapause development was lower than the temperature threshold of larvae in continuous development. This is important for adjusting phenological models in S. nonagrioides and in other species, and may explain why in many cases adults appear in the field when the supposed temperature threshold for development has not been attained.     

Origin and taxonomic status of the Palearctic population of the stem borer Sesamia nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae)

The major pest of maize in Mediterranean Europe, the stem borer Sesamia nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae), has a fragmented distribution, north and south of the Sahara. The present study aimed: (1) to clarify the uncertain taxonomic status of the Palearctic and sub‐Saharan populations which were first considered as different species and later on as subspecies (Sesamia nonagrioides nonagrioides and Sesamia nonagrioides botanephaga) and (2) to investigate the origin of the Palearctic population which extends from Spain to Iran, outside what is considered typical for this mainly tropical genus. We reconstructed the evolutionary history of both populations using one nuclear and two mitochondrial genes. The sub‐Saharan taxon was fragmented in two isolated populations (West and East) whose mitochondrial genes were distant by 2.3%. The Palearctic population was included in the East African clade and its genes were close or identical to those of a population from Central Ethiopia, where the species was discovered for the first time. Similarly, in Africa, the alleles of the nuclear gene were distributed mainly in two West and East clades, whereas some Palearctic alleles belonged to the West clade. The Palearctic population originated therefore from East and West Africa and is the progeny of the cross between these two African populations. The main species concepts were in agreement, leading to the conclusion that the three populations are still conspecific. In the surveyed regions, the species therefore does not include two subspecies but three isolated populations. The Palearctic population suffered from severe bottlenecks that resulted in the fixation of one East African mitochondrial genome and the large reduction in its genetic diversity compared to the African populations. The data suggest that natural colonization of the Palearctic region was more plausible than human introduction. The allelic distribution of the Palearctic population was similar to that of species that survived the last glaciation. It is concluded that the African populations expanded during the last interglacial, crossed the Sahara and mixed in North Africa where fixation of the East mitochondrial genome occurred. The species then colonized Europe westward through only one eastern entrance. The coalescent‐based estimate of the time to the ancestor of the Palearctic population was 108 000 years, which is consistent with this scenario. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 904–922.     

Susceptibility to the Cry1F toxin of field populations of Sesamia nonagrioides (Lepidoptera: Noctuidae) in Mediterranean maize cultivation regions

Maize hybrids expressing the Cry1F toxin provide efficient control of lepidopteran pests. The Mediterranean corn borer, Sesamia nonagrioides (Lefèvre), is one of the most damaging pests of maize in the Mediterranean basin. In this work we firstly determined the efficacy of maize hybrids expressing the Cry1F toxin (event TC1507) to control neonates of S. nonagrioides. Leaf tissue feeding bioassays revealed that TC1507 maize is highly effective against this pest, and the percentage mortality obtained was comparable to that obtained with a Cry1Ab-expressing maize hybrid (Compa CB, event 176), which is known to be highly efficacious against S. nonagrioides. Secondly, interpopulation variation in the susceptibility to the Cry1F insecticidal protein was established for nine field-collected populations of S. nonagrioides (three Spanish, two French, two Italian, one Greek, and one Turkish). Estimates of the susceptibility of larvae to the Cry1F toxin showed low variability in lethal concentrations and growth inhibition concentrations among field populations. Moreover, no significant differences were found when they were grouped by geographical areas [Western Mediterranean (Spain and France) versus Eastern Mediterranean (Italy, Greece and Turkey)] or by history of exposure to Bt plants (Spanish vs. other populations). Therefore, the minor differences found in field populations can be attributed to natural variation in sensitivity to Cry1F. The importance of establishing baselines of susceptibility for resistance detection is discussed. Future changes in susceptibility of S. nonagrioides populations to Cry1F could be documented based on this baseline data.     

Effect of DIMBOA on growth and digestive physiology of Sesamia nonagrioides (Lepidoptera: noctuidae) larvae

The hydroxamic acid 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) was assessed for its effect on growth and digestive physiology of larvae of the stalk corn borer Sesamia nonagrioides Lef. Nutritional indices and activities of some digestive and detoxification enzymes were determined for larvae feeding on a DIMBOA-containing diet for the first two days of the third instar (short-term feeding assays), and from neonates to third instar (long-term feeding assays). DIMBOA reduced the relative growth rate and the efficiency of conversion of ingested food without affecting the relative consumption rate in long-term feeding assays, but it had no effect in short-term assays. Moreover, elastase-like activity was significantly increased by DIMBOA in short-term feeding assays, whereas microsomal oxidase activity was increased and esterase activity was reduced in long-term feeding assays. In vitro, DIMBOA inhibited the activities of carboxypeptidases, aminopeptidase, glutathione S-transferase and esterase, but it had no effect on trypsin, chymotrypsin and elastase. The implications of the altered levels of proteases and detoxification enzyme activities on the digestive physiology of larvae feeding on DIMBOA-containing diets are discussed.     

Mitochondrial DNA Restriction Map and Cytochrome c Oxidase Subunits I and II Sequence Divergence of Corn Stalk Borer Sesamia nonagrioides (Lepidoptera: Noctuidae)

Corn stalk borer Sesamia nonagrioides (Lefebvre) (Lepidoptera: Noctuidae) is among the most important insect pests of corn in the Mediterranean basin. The mitochondrial DNA of this insect was purified and a restriction map was constructed. The size of the mtDNA genome is 16.3 kb. Genetic analysis of four corn stalk borer populations, collected from Greece (three populations) and Spain (one population), was undertaken using DNA sequences of the mtDNA cytochrome oxidase (CO) I and II genes. Sequencing of a 2079 bp region of these genes revealed 25 polymorphic sites among the populations. Five molecular RFLP markers, located in the mtDNA COI and COII genes, were surveyed, and two different haplotypes were detected. Phylogenetic analysis based on COI/COII nucleotide sequences revealed genetic differentiation between samples, and the results are discussed in relation to the geographic distribution of the corn stalk borer in two Mediterranean countries.